The home side are currently top of Group H with 10 points, while Valencia sit second, level on eight points with third-placed Chelsea, who complete their group-stage campaign against Lille.
Top of the section on 10 points, Ajax will head into Tuesday's match knowing that a draw would be enough for them to book their spot in the next round of the competition. That said, the Dutch giants would also qualify with a defeat if Chelsea fail to overcome Lille in the section's other match.
Ajax have won three, drawn one and lost one of their five group-stage matches this term, with their only dropped points coming against Chelsea. Indeed, they suffered a 1-0 defeat at home to the Blues before claiming a point in a 4-4 draw at Stamford Bridge at the start of last month.
The 2-0 win at Lille on November 27 was a huge result for Erik ten Hag's side, who made the semi-finals of last season's Champions League before losing to Tottenham Hotspur in spectacular fashion.
Incredibly, Ajax have actually lost six of their last eight home matches against La Liga opposition, although they played out a 1-1 draw with Valencia when Los Che last visited Amsterdam during the 2002-03 competition.
Ten Hag's side will actually enter this match off the back of a 2-0 home loss to Willem II in the league on Friday night, although a strong start to the 2019-20 campaign has left them top of the Eredivisie.
Valencia will book their spot in the next round of the competition if they manage to beat Ajax on Tuesday night, or providing that they finish at least level on points with third-placed Chelsea.
The English giants enter their clash with Lille knowing that a win would be enough to advance, meanwhile, meaning that it is very much a three-way battle to secure the two qualification spots in Group H.
Los Che made a perfect start to their group-stage campaign as they recorded a 1-0 win at Chelsea, but they have drawn two of their last three in the competition, including a 2-2 at home to Chelsea last time out.
The Spanish giants have not actually reached the last-16 stage of the Champions League since 2012-13, but they have history in the competition having made the final in both 2000 and 2001.
What's more, Valencia have lost just two of their last 13 matches against Dutch opposition, while they will enter this game off the back of successive La Liga victories, including an impressive 4-2 success away to Levante on Saturday night.
Ajax form (Champions League): WWLDW
Ajax form (all competitions): DWWWWL
Valencia form (Champions League): WLDWD
Valencia form (all competitions): WWLDWW
Quincy Promes is also a doubt having missed out against Willem II on Friday with a muscular problem, but Ten Haag has no fresh injury concerns from last week's Eredivise clash.
The fact that the Dutch side have issues in the final third could see experienced forward Klaas-Jan Huntelaar start, while there will also be a spot for Hakim Ziyech, who has incredibly provided 20 assists in just 25 appearances at club level during the 2019-20 campaign.
As for Valencia, Cristiano Piccini, Goncalo Guedes, Denis Cheryshev and Geoffrey Kondogbia remain on the treatment table, but Jaume Costa and Ezequiel Garay both made their returns from injury at the weekend.
The visitors could be without number one goalkeeper Jasper Cillessen, though, with the Netherlands international in a race to recover from the injury that he picked up against Levante.
Ajax possible starting lineup: Onana; Veltman, Schuurs, Blind, Tagliafico; Van de Beek, Mazraoui; Tadic, Ziyech, Lang; Huntelaar
Valencia possible starting lineup: Domenech; Costa, Garay, Gabriel, Gaya; Soler, Coquelin, Parejo, Torres; Gameiro, Rodrigo
We say: Ajax 1-1 Valencia
We fancy Ajax to get the point that they need to book their spot in the next round, and it will then be a case of wait and see for Valencia as they hope for a favour from Lille against Chelsea.